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    RBC

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    RBC

    Uploaded by

    aditya32157
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    RBC

    Uploaded by

    aditya32157

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    © All Rights Reserved
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    of 6

    Experiment 6

    To Determine the Red Blood Cell (RBC)


    Count of Own Blood Sample
    Aim:
    To determine the Red Blood Cellcount of own blood sample.
    Requirements:
    Haemocytometer set consisting of Neubauers counting chamber and a RBC dilution
    pipette
    Compound microscope
    Pricking needle
    Absorbent cotton
    70% alcohol
    " Coverslip
    -Rubber tube

    101
    -Red bead

    0.5
    Stem

    Fig.6.1: RBC diluting pipette


    Properly Counting pntues
    loaded chambers Flooded

    Cover Aif NUnder


    glass 2 bubbles 9i90nsiC loaded
    (a) Neubauer chamber (b) Chamber loaded with blood sample

    (6.1)
    Human Anatomy and Physiology (Practical) 6.2 To Determine the Red Blood Cell

    WBC WBC

    Teneups
    mooomeaH
    WBC WBC
    omouogmoD

    00neuioedA
    Fig. 6.3: Enlarged viewof Neubauers countingchamber0

    -RBCcells
    deseavv,

    Fig. 6.4: Zig-Zag counting of RBC heqons


    aedmer bebeol
    Theory:Red Blood Cells have:
    Shape: Circular, biconcave,non-nucleated disc like structure.
    Salient feature: Contain haemoglobin.
    Count: 5.4 million RBCS per microlitre (ul) of blood in male and 4.8 million RBCs per
    ul of blood in female.
    Size: Diameter 7-8 um and Thickness 2 um.
    Function:ubsbeol sodst (d
    o Transportation of oxygen and carbon dioxide, maintenance of acid base balance,
    ionic balance and blood viscosity.
    Human Anatomy and Physiology (Practical) 6.3 To Determine the Red Blood Cell...
    RBCs contain larger surface area because of biconcavity for oxygen diffusion.
    They can easily squeeze through the capillaries.
    The formation of red blood occurs in the red bone marrow, the process is called as
    erythropoiesis.
    " During fetal life erythropoiesis occurs in the spleen, liver, thymus and bone marrow.
    After birth, it is confined to the red bone marrow.

    2ouh Fig. 6.5: Red blood cels6bd hss D83 sdT


    Clinical Significance:
    The total RBC count is performed to access the red cell mass in the blood.
    The change in erythrocyte number is detected in clinical practice by ordering the
    estimation of haemoglobin rather than total RBC count as estimation of
    haemoglobin is easy and less expensive.
    Conditions that Decrease the RBC Count:
    Pregnancy due to blood dilutionta iie2Oeaholrb nubo?
    Different types of anaemiaUavghocS)5isdgluemuibo2
    Dazua0)sbnoiusihubyshM
    Bone marrow f a i l u r e h
    aviees ba alo0iwbalieiC
    Internal or external bleeding
    L Leukemia
    Malnutrition
    Thyroid disorders
    " Nutritional deficiencies of iron, copper, folate, and vitamins Bs and B12
    Conditions that Increase the RBC Count:
    New born have high count
    eogdinsi
    Excessive sweating
    Human Anatomy and Physiology (Practical) 6.4 To Determine the Red Blood Cell
    Severe diarrhoea and vomiting causes severe loss of fluid due to hemoconcentration
    " Cigarette smoking
    Congenital heart disease 2lasiogoris
    pnug
    " Renal cell carcinoma (kidney cancer)esictoh9b9
    Pulmonary fibrosis
    RBCS)
    Polycythemia vera (Bone marrow disease that causes overproduction of
    Principle:
    red blood cells is too high and size is too small. Therefore, it is not
    Tne number of
    possible to count the number of RBCs under high power.
    dilution of blood with RBC diluting fluid to
    Ihis problem of counting is overcome by
    a known amount.
    Neauber chamber and the cells are
    Then, the diluted bloodsample is loaded on the
    allowed to settle and counted under the microscope (40 X).
    with dilution factor and volume
    The RBC can be calculated by multiplying it
    correction factor.
    whole blood is calculated.
    The number of red blood cells per mm of undiluted
    Composition of RBC Diluting Fluid:lb at tadhm n elppins ci acsro sri
    fluid is used as an
    o The most frequently used fluid is Hayem's fluid and Dacie's
    alternative to Hayem's fluid. avenge el bnn veseainidolpomssr
    1. Hayem's fluid:
    o Sodium chloride (0.5 g):It maintains osmolarity.old of eub vonsnpor9
    o Sodium sulphate (2.5 g): It prevents aggregation of RBCs.nohd
    o Mercuric chloride (0.25 g): It is used as a preservative.
    o Distilled water (100ml): It is used as a solvent.
    2. Dacie's fluid:
    o Trisodium citrate (3.13 g): It prevents aggregation of RBCs and provide
    correct osmotic pressure. noithtunlaM
    o Formaldehyde (1 ml): It isused as preservative.
    Distilled water (100 ml): It is used as a solvent.
    Procedure:
    Clean and dry the RBC diluting pipette, watch glass, coverslip and Neubauers
    chamber properly.
    Take enough RBC diluting fluid in the watch glass.
    (Practical) sotba To Determine the Red Blood Cell ...
    Human Anatomy and Physiology
    6.5

    Sterilize the finger with 70% alcohol and prick it under aseptic condition.
    the
    .Touch the tip of RBC diluting pipette to blood drop and suck the blood upto
    0.5 mark.
    Again dip the tip of RBC diluting pipette into the watch glass to suck the RBC
    diluting fluid exactly upto the mark 101.
    Then hold the pipette horizontally and shake for 10 seconds for proper mixing of
    blood with the RBC diluting fluid.
    stem does not
    Discard the first two drops of fluid from the pipette as the fluid in the
    contain RBC.
    blood and allow to stand for two
    Then charge the Neubauers chamber with diluted
    minutes as the cels get settles down.
    microscope and adjust the
    B Place the charged Neubauers chamber on the stage of
    microscope under high power objective (40 X).
    Count the total number of RBCs in the five squares.
    cells present in the square and
    Toavoid duplicate counting of cells again,count thecount the cells on its right and
    not
    those present in the left and lower lines. Do
    upper lines.
    five chambers.
    "Finally calculate the total number of RBC in
    R= R1 + Rg+ R3 + R4 + Rs
    Calculation:
    1. Dilution correction factor:
    We have diluted 0.5 parts of blood in 100
    parts of solution (Blood + RBC diluting
    fluid).
    So dilution is: 0.5 to100 means, 1 to 200
    200.
    " So, the dilution correction factor is
    2. Volume correction factor:
    " Each Rsquare has an area of 0.04 sq.
    mm and depth of 0.1 mm.
    Rsection.
    Volume of 1 Rsection = Areaof 1 Rsection x Depth of 1
    0.1 cu. mm = 0.04 sg. mm x 0.1 mm
    Total volume of 5 Rsections = Volume of 1 Rsection x 5
    = 0.04 x5
    =0.02 cubic millimetre (cu. mm)
    " Volume correction factor = Volume desired/Volume used
    1.0
    Volume correction factor = 0.02= 50
    Human Anatomy and Physiology (Practical) 6.6 (sst To Determine the Red Blood Cell ..

    " Fornmula for RBC count = Total number of RBC] [Volume correctionDilution
    in 5 R sections factor factor
    en
    Result: The red blood cells count of own blood sample was found to be
    -----/cu. mm of blood.
    Report:
    Vono Patient Name:
    Sample Number:
    1on 25 o Observed RBC count: ina
    Normal RBC count:
    ow 10o Method used:s boold befuib niyemsno 219uscuslf srt
    Date:
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